Exposing synthetic lethal vulnerabilities in EBV-positive AIDS-NHL through novel replication dependency factors

通过新型复制依赖性因子揭示 EBV 阳性 AIDS-NHL 的综合致命脆弱性

• 批准号: 10700376
• 负责人: SUMITA BHADURI-MCINTOSH
• 金额: $ 61.18万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-04 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10700376
• 关键词: AIDS-Related Lymphoma; Acquired Immunodeficiency Syndrome; Address; B-Cell Activation; B-Cell Lymphomas; B-Lymphocytes; Biopsy; Blast Cell; Blood; Burkitt Lymphoma; Cancer Etiology; Cell Cycle; Cell Death; Cell Line; Cell Nucleus; Cell Proliferation; Cells; Cessation of life; Complex; DNA; DNA biosynthesis; DNA replication fork; Dependence; Development; Disease; Ensure; Epstein Barr Virus lymphoma cell; Epstein-Barr Virus-Related Lymphoma; Event; Genes; Genetic; Genome; Goals; HIV; HIV/AIDS; Human Herpesvirus 4; Immune; Immunocompromised Host; Immunologic Surveillance; Impairment; Individual; Infection; Influentials; Investigation; Job' s Syndrome; Knowledge; Link; Lymphoma; Lymphoma cell; MCM7 gene; Malignant Neoplasms; Mass Spectrum Analysis; Mechanics; Modeling; Morbidity - disease rate; Mutation; Non-Hodgkin' s Lymphoma; Nucleotides; Oncoproteins; Pathway interactions; Patients; Persons; Phase; Predisposition; Proliferating; Property; Proteins; Proteome; Proto-Oncogenes; Public Health; Risk; Role; S phase; STAT3 gene; Stress; Testing; Therapeutic; Tonsil; Trans-Activators; Transcript; Translating; Transplant Recipients; Up-Regulation; Viral; Virus; Virus Latency; Zinc Fingers; antiretroviral therapy; cancer cell; cancer specimen resource; cell transformation; chemotherapy; clinically relevant; experience; gammaherpesvirus; genome-wide; helicase; immunosuppressed; improved outcome; in vivo; infected B cell; innovation; large cell Diffuse non-Hodgkin' s lymphoma; lymphoblastoid cell line; mortality; new therapeutic target; novel; oral pathogen; recruit; replication stress; transforming virus; tumor microenvironment

PROJECT SUMMARY Virus-associated lymphomas cause significant morbidity and mortality in HIV-infected individuals – indeed, the oral pathogen Epstein-Barr virus (EBV) contributes to up to 90% of diffuse large B-cell lymphomas (DLBCL) and 40% of Burkitt lymphomas (BL). Although combined antiretroviral therapy (cART) and chemotherapy have improved outcomes for AIDS lymphomas, challenges remain particularly with virus-associated AIDS lymphomas, prompting efforts to better understand virus-associated factors and pathways. In particular, EBV-driven cellular genome replication which is essential to lymphoma proliferation remains underexplored. Upon infection of B cells, EBV drives host DNA replication which is essential for establishment of viral latency as well as proliferation of cancer cells. However, such viral oncoprotein-driven DNA replication is plagued with physical and functional obstacles, resulting in replication stress. Such replication stress is a barrier to cancer. And yet, how EBV-cancer cells overcome such stress at replication forks to successfully proliferate is not well understood. In addressing this knowledge gap, we combined isolation of proteins on nascent DNA (iPOND) and mass spectrometry to discover novel fork proteins. This revealed a critical role for ZC3H18 (or ZC3) as a replication dependency factor that EBV upregulates to ensure host genome replication and lymphoma cell proliferation; notably, ZC3 had not been previously linked to DNA replication. Indeed, EBV+ DLBCL from AIDS patients have elevated ZC3 expression compared to EBV- lymphomas. An intrinsically disordered protein, ZC3 has the potential to concentrate a variety of proteins at replication forks. We find a direct interaction between ZC3 and MCM7 (a core component of the replicative helicase complex), further pointing to ZC3’s influential role in proliferation of EBV transformed cells. Importantly, ZC3’s partnership with other replication dependency factors exposes EBV-lymphoma cells to synthetic lethality – such therapies exploit the property that cancer cells tolerate perturbation of a single gene but succumb to co-disruption of multiple genetic events. In this application, we will test the hypothesis that EBV modulates the DNA replication machinery, ensuring proliferation of transformed cells in the face of replication stress and enhancing the potential for susceptibility to synthetic lethality. We will perform the following aims using ex vivo models and translate our results to patient- derived EBV+ AIDS lymphomas obtained from the AIDS and Cancer Specimen Resource (ACSR). Aim 1. Investigate how novel dependency factors unmask synthetic lethal vulnerabilities in EBV- transformed cells & Aim 2. Investigate mechanisms of ZC3 upregulation, replication machinery rewiring, and contribution of replication dependency factors to EBV+ AIDS lymphomas. These studies will identify mechanisms and generate new paradigms that reveal how an opportunistic virus modulates the host replication machinery to maintain the transformed state. Our long-term goal is to identify novel druggable targets that demonstrate synthetic lethality against EBV+ lymphomas in persons with HIV/AIDS.

项目摘要 病毒相关的淋巴瘤在HIV感染的个体中引起明显的发病率和死亡率 - 实际上， 口腔病原体爱泼斯坦 - 巴尔病毒（EBV）贡献了多达90％的弥漫性大B细胞淋巴瘤（DLBCL）和 伯基特淋巴瘤（BL）的40％。尽管抗逆转录病毒疗法（CART）和化学疗法的联合结合 改善了艾滋病淋巴瘤的结果，尤其是与病毒相关的艾滋病淋巴瘤的挑战， 促使努力更好地了解病毒相关的因素和途径。特别是，EBV驱动的蜂窝 对于淋巴瘤增殖至关重要的基因组复制仍然没有被逐渐倍增。 在感染B细胞后，EBV驱动宿主DNA复制，这对于建立病毒潜伏期至关重要 以及癌细胞的增殖。但是，这种病毒癌蛋白驱动的DNA复制困扰着 物理和功能障碍，导致复制应力。这种复制应力是癌症的障碍。 然而，在复制叉上如何克服这种压力以成功增殖的情况不好 理解。在解决这一知识差距时，我们将蛋白质的分离在新生的DNA（IPOND）和 质谱法以发现新型的叉蛋白。这揭示了ZC3H18（或ZC3）的关键作用 EBV上调以确保宿主基因组复制和淋巴瘤细胞的复制依赖性因素 增殖;值得注意的是，ZC3以前没有与DNA复制相关。确实，来自艾滋病的ebv+ dlbcl 与EBV淋巴瘤相比，患者的ZC3表达升高。本质上无序的蛋白质ZC3 有可能在复制叉时集中多种蛋白质。我们发现 ZC3和MCM7（复制性解旋酶复​​合酶的核心组成部分），进一步指出了ZC3的影响力 在EBV转化细胞的增殖中。重要的是，ZC3与其他复制依赖性因素的合作关系 将EBV淋巴瘤细胞暴露于合成致死性 - 这种疗法利用了癌细胞耐受性的特性 单个基因的扰动，但屈服于多个遗传事件的共同消失。 在此应用程序中，我们将测试EBV调节DNA复制机制的假设，以确保 面对复制应力时转化细胞的增殖，并增强了对易感性的潜力 综合致死性。我们将使用离体模型执行以下目标，并将结果转化为患者 衍生的EBV+ AIDS淋巴瘤从艾滋病和癌症标本资源（ACSR）获得。 目标1。研究新的依赖性因素如何揭示EBV-中的合成致命性脆弱性 转化的细胞和目标2。研究ZC3上调的机制，复制机制重新布线， 复制依赖因子对EBV+ AIDS淋巴瘤的贡献。 这些研究将确定机制并产生新的范式，以揭示机会病毒如何 调节主机复制机制以维护转换状态。我们的长期目标是确定 具有HIV/AIDS患者的EBV+淋巴瘤的合成性致死性的新型可药物靶标。